Composition for preventing and / or treating diabetic cardiomyopathy and application thereof

Abstract

The invention relates to the technical field of medicines, in particular to a composition for preventing and / or treating diabetic cardiomyopathy and application thereof. The invention provides a composition for preventing and / or treating diabetic cardiomyopathy. The composition comprises the following components: spermidine, coenzyme Q, notoginsenoside and resveratrol, the mass ratio of the spermidine to the coenzyme Q to the notoginsenoside to the resveratrol is (0.3-1): (6-20): (3-15): (0.1-0.5). According to the composition, four active ingredients including spermidine, coenzyme Q, notoginsenoside and resveratrol are scientifically matched, so that the composition shows a remarkable synergistic interaction effect and a multiple protection mechanism in treatment of diabetic cardiomyopathy; moreover, the composition has the characteristic of high biological safety, is easy to absorb and metabolize by a human body, and has small side effects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for preventing and / or treating diabetic cardiomyopathy and applications thereof. Background Art

[0002] Diabetes, a chronic metabolic disease characterized by endocrine and metabolic disorders, is becoming increasingly prevalent worldwide. Diabetes is a significant risk factor for ventricular remodeling, promoting myocardial pathological processes such as myocardial fibrosis, apoptosis, and hypertrophy through multiple mechanisms, including metabolic disorders, oxidative stress, and inflammation. Early manifestations include decreased diastolic function, which progresses to heart failure with reduced ejection fraction, ultimately leading to severe heart failure.

[0003] Diabetic ventricular remodeling is a complex pathological process driven by multiple factors, requiring the integration of metabolic regulation, anti-inflammatory, and anti-fibrotic strategies. Currently, traditional drug treatments for diabetic cardiac function impairment (especially with ventricular remodeling) mainly include RAAS inhibitors, β-blockers, diuretics, and aldosterone receptor antagonists. Although these drugs are effective in general heart failure patients, they often face limitations in diabetic patients. RAAS inhibitors often damage myocardial metabolism in diabetic patients due to worsening insulin resistance or persistent hyperglycemia, and have side effects such as hyperkalemia and worsening renal function. β-blockers may worsen insulin resistance and lipid metabolism disorders and have limited effects on diastolic dysfunction. Diuretics are prone to hypokalemia and hyperuricemia, aggravating insulin resistance and the risk of arrhythmias, and are unable to reverse ventricular remodeling. Aldosterone receptor antagonists are limited by renal function and are ineffective against microvascular lesions. Therefore, finding new, effective treatments with fewer side effects is urgently needed. Summary of the Invention

[0004] The object of the present invention is to provide a composition for preventing and / or treating diabetic cardiomyopathy, which exhibits significant synergistic effects and multiple protective mechanisms in the treatment of diabetic cardiomyopathy; and the composition has high biosafety characteristics, is easily absorbed and metabolized by the human body, and has few side effects.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a composition for preventing and / or treating diabetic cardiomyopathy, comprising the following components: spermidine, coenzyme Q, notoginseng saponin and resveratrol;

[0007] The mass ratio of spermidine, coenzyme Q, notoginseng saponin and resveratrol is 0.3-1:6-20:3-15:0.1-0.5.

[0008] Preferably, the mass ratio of spermidine, coenzyme Q, notoginseng saponin and resveratrol is 0.5-0.8:10-16:6-12:0.2-0.4.

[0009] The present invention also provides the use of the composition in preparing a preparation for preventing and / or treating diabetic cardiomyopathy.

[0010] The present invention also provides a preparation for preventing and / or treating diabetic cardiomyopathy, comprising the composition and pharmaceutically acceptable excipients.

[0011] Preferably, the excipients include one or more of fillers, binders, disintegrants, lubricants, solvents and sustained-release materials.

[0012] Preferably, the preparation is in the form of tablets, capsules, granules, injections or oral solutions.

[0013] Beneficial effects of the present invention:

[0014] The invention provides a composition for preventing and / or treating diabetic cardiomyopathy. By scientifically combining four active ingredients: spermidine, coenzyme Q, notoginseng saponins, and resveratrol, the composition exhibits significant synergistic effects and multiple protective mechanisms in the treatment of diabetic cardiomyopathy. Furthermore, the composition has high biosafety, is easily absorbed and metabolized by the human body, and has minimal side effects.

[0015] This combination, through the synergistic effects of multiple components, multiple targets, and multiple pathways, overcomes the single-target limitations of existing diabetic cardiomyopathy treatments, demonstrating significant advantages in improving cardiac function, inhibiting inflammatory responses, and reversing myocardial fibrosis. Its unique formulation enhances the bioavailability of its components, significantly reducing dosage and potential side effects while maintaining efficacy. This combination offers new insights into the clinical treatment of diabetic cardiomyopathy and holds significant clinical and practical value. DETAILED DESCRIPTION

[0016] The present invention provides a composition for preventing and / or treating diabetic cardiomyopathy, comprising the following components: spermidine, coenzyme Q, notoginseng saponin and resveratrol;

[0017] The mass ratio of spermidine, coenzyme Q, notoginseng saponin and resveratrol is 0.3-1:6-20:3-15:0.1-0.5, preferably 0.5-0.8:10-16:6-12:0.2-0.4, and more preferably 0.65:13:9:0.3.

[0018] In the present invention, spermidine is preferably naturally extracted spermidine with a purity of ≥98%. Naturally extracted spermidine has better biocompatibility and higher safety.

[0019] Coenzyme Q is preferably coenzyme Q10 with a purity of ≥99%. Coenzyme Q10 plays an important role in human energy metabolism. High-purity coenzyme Q10 can better ensure its biological activity.

[0020] The purity of notoginsenosides is ≥98%;

[0021] The purity of resveratrol is ≥98%. Resveratrol has multiple biological activities such as antioxidant, anti-inflammatory, and cardiovascular protection. It can synergize with other components to further enhance the therapeutic effect of the composition.

[0022] In the present invention, spermidine can activate AMPK to initiate autophagy, resveratrol activates SIRT1 to enhance deacetylation, and coenzyme Q has an antioxidant effect and can protect the mitochondrial membrane. The three together enhance myocardial antioxidant capacity and reduce MDA content. In addition, coenzyme Q can enhance the electron transfer capacity of the mitochondrial respiratory chain, promote ATP production, and increase cell vitality. When combined with spermidine, it can better maintain cell homeostasis and improve myocardial energy metabolism.

[0023] Spermidine can inhibit the activation of inflammatory signaling pathways such as nuclear factor-κB (NF-κB) and reduce the release of inflammatory factors such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6); Panax notoginseng saponins can downregulate the expression of inflammatory mediators and reduce the infiltration of inflammatory cells in myocardial tissue. The two synergistically exert anti-inflammatory effects, effectively reducing the degree of inflammatory response in myocardial tissue; resveratrol can enhance the anti-inflammatory effect by inhibiting the activity of inflammatory-related enzymes. Working together with spermidine and Panax notoginseng saponins, the combination shows a better effect in reducing TNF-α and IL-6 levels.

[0024] Panax notoginseng saponins have the effect of promoting blood circulation and removing blood stasis. They can dilate coronary arteries, increase myocardial blood flow, improve myocardial microcirculation, and provide sufficient oxygen and nutrients to myocardial cells. At the same time, they can also inhibit platelet aggregation, prevent thrombosis, and avoid further aggravation of myocardial ischemia. Spermidine can inhibit the activation and proliferation of myocardial fibroblasts and reduce collagen synthesis. Resveratrol can downregulate the expression of fibrosis-related proteins such as type I collagen and type III collagen. The three work synergistically to effectively improve the degree of myocardial fibrosis.

[0025] Therefore, spermidine, coenzyme Q, notoginseng saponins and resveratrol form a mutually coordinated and complementary treatment system through synergistic effects in multiple aspects such as anti-oxidation, anti-inflammation, improving myocardial metabolism and microcirculation, and inhibiting fibrosis, which significantly enhances the therapeutic effect on diabetic cardiomyopathy.

[0026] The present invention also provides the use of the composition in preparing a preparation for preventing and / or treating diabetic cardiomyopathy.

[0027] The present invention also provides a preparation for preventing and / or treating diabetic cardiomyopathy, comprising the composition and pharmaceutically acceptable excipients.

[0028] In the present invention, the excipients include one or more of fillers, binders, disintegrants, lubricants, solvents and sustained-release materials;

[0029] The filler includes one or more of starch, lactose, mannitol and microcrystalline cellulose.

[0030] The binder includes hypromellose and / or povidone;

[0031] The disintegrant includes sodium starch glycolate and / or cross-linked polyvinylpyrrolidone;

[0032] The lubricant includes magnesium stearate and / or talc;

[0033] The solvent includes water or ethanol;

[0034] The sustained-release material includes one or more of hypromellose K4M, hypromellose K15M and ethyl cellulose.

[0035] In the present invention, the dosage form of the preparation is tablet, capsule, granule, injection or oral solution.

[0036] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1 A composition for preventing and / or treating diabetic cardiomyopathy

[0038] Spermidine (purity ≥ 98%), coenzyme Q10 (purity ≥ 99%), notoginseng saponin (purity ≥ 98%) and resveratrol (purity ≥ 98%) were mixed at a mass ratio of 0.5:15:10:0.2.

[0039] Example 2 A composition for preventing and / or treating diabetic cardiomyopathy

[0040] Spermidine (purity ≥98%), coenzyme Q10 (purity ≥99%), notoginseng saponin (purity ≥98%) and resveratrol (purity ≥98%) were mixed at a mass ratio of 0.3:18:5:0.1.

[0041] Example 3 A composition for preventing and / or treating diabetic cardiomyopathy

[0042] Spermidine (purity ≥98%), coenzyme Q10 (purity ≥99%), notoginseng saponin (purity ≥98%) and resveratrol (purity ≥98%) were mixed at a mass ratio of 1:8:12:0.3.

[0043] Comparative Example 1: A composition for preventing and / or treating diabetic cardiomyopathy

[0044] Coenzyme Q10 (purity ≥ 99%), notoginseng saponin (purity ≥ 98%) and resveratrol (purity ≥ 98%) were mixed at a mass ratio of 15:10:0.2.

[0045] Comparative Example 2: A composition for preventing and / or treating diabetic cardiomyopathy

[0046] Spermidine (purity ≥98%), coenzyme Q10 (purity ≥99%) and notoginseng saponin (purity ≥98%) were mixed at a mass ratio of 0.5:15:10.

[0047] Experimental Example 1

[0048] Construction of animal model:

[0049] Model Construction: SD rats (6-8 week old Sprague-Dawley rats, weighing 180-200 g, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) were intraperitoneally injected with streptozotocin (STZ) in citrate buffer (pH 4.5) at a dose of 60 mg / kg for 5 consecutive days. Blood glucose levels were monitored starting 72 hours after injection. Tail vein blood glucose levels were measured in rats fasting for 12 hours for 3 consecutive days. Successful modeling was determined when blood glucose levels remained above 16.7 mmol / L, thus establishing a type 1 diabetic rat model (DM model).

[0050] Treatment group (n=6): DM model rats were intraperitoneally administered with the compositions described in Example 1 and Comparative Examples 1-2 (dosage: 5 mg / kg) daily for 8 consecutive weeks;

[0051] Model group (n=6) (DM group): DM model rats were used as positive control, and the injection method of the treatment group was referred to, and the same volume of normal saline was injected as that of the treatment group;

[0052] Control group (n=6) (Ctrl group): Normal SD rats were used as negative control group, and the injection method was referred to that of the treatment group, and the same volume of normal saline was injected as that of the treatment group;

[0053] The body weight and blood glucose levels of the rats in each group were recorded weekly, and the results are shown in Table 1. The left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), left ventricular internal diameter at end-diastole (LVIDd), and left ventricular internal diameter at end-systole (LVIDs) of the rats were measured using a Philips IE33 ultrasound system under isoflurane-induced anesthesia (2% concentration), and the results are shown in Table 2.

[0054] Table 1 Body weight and blood glucose measurement results of rats

[0055]

[0056] As can be seen from Table 1, 4 weeks after injection of the composition described in Example 1, the body weight of the rats increased significantly compared with the DM group, and the blood sugar level decreased significantly (P<0.05).

[0057] Table 2 Rat heart evaluation results

[0058] Group LVEF (%) LVFS (%) LVIDd (mm) LVIDs (mm) Control group 78.5±3.2 42.1±2.1 7.2±0.3 3.8±0.1 DM group 52.3±4.5 24.6±2.8 8.6±0.4 6.4±0.3 Example 1 68.4±3.8 36.2±2.5 7.8±0.3 4.8±0.2 Comparative Example 1 58.1±4.0 28.7±2.3 8.2±0.4 5.6±0.4 Comparative Example 2 62.6±3.6 32.5±2.4 8.1±0.2 5.3±0.2

[0059] As can be seen from Table 2, compared with the DM group, the LVEF and LVFS of the Example 1 group, the Comparative Example 1 group, and the Comparative Example 2 group were significantly increased, and the LVIDd and LVIDs were significantly decreased (P<0.05); the cardiac function indicators of the Example 1 group were most significantly improved, and the differences were significant compared with the Comparative Example 1 group and the Comparative Example 2 group (P<0.05); therefore, the composition described in Example 1 can effectively improve the left ventricular ejection and shortening capacity, and reduce the ventricular cavity diameter.

[0060] Experimental Example 2: Determination of ventricular fibrosis

[0061] Rats were treated by overdose of anesthesia and killed. The heart was quickly removed and residual blood was flushed with normal saline. The left ventricular myocardial tissue was obtained, one part of which was fixed in 4% paraformaldehyde and the other part was stored in a -80°C ultra-low temperature refrigerator for future use.

[0062] Myocardial tissue stored at -80°C was prepared and RIPA lysis buffer (containing protease inhibitors) was added at a ratio of tissue weight to lysis buffer of 1:10. The tissue was homogenized on ice and centrifuged at 4°C (12,000 rpm, 15 min). Total protein was extracted and the expression of fibrosis-related proteins (type I collagen and α-SMA protein) was detected by Western blotting. The results are shown in Table 3.

[0063] Specific steps for detecting fibrosis-related proteins by western blotting:

[0064] (1) Protein concentration determination: The total protein concentration of the extracted protein was determined using a BCA protein quantification kit. According to the kit instructions, the standard was diluted in a gradient manner. Then, appropriate amounts of the standard and the protein sample to be tested were added to a 96-well plate. BCA working solution was then added. After incubation at 37°C for 30 minutes, the absorbance was measured at a wavelength of 562 nm using a microplate reader. The concentration of the protein sample to be tested was calculated based on the standard curve.

[0065] (2) Protein denaturation: Based on the measured protein concentration, take an appropriate amount of protein sample and add 5× SDS loading buffer to make the final concentration of the loading buffer 1×. After thorough mixing, boil the sample in boiling water for 10 min to denature the protein, and then cool it on ice.

[0066] (3) SDS-PAGE electrophoresis: Prepare 10% separating gel and 5% stacking gel; load 30 μg of denatured protein sample per well, along with a protein molecular weight standard; perform electrophoresis at 80 V. Once the protein sample enters the separating gel, adjust the voltage to 120 V and continue electrophoresis until the bromophenol blue indicator reaches the bottom of the gel, then stop electrophoresis;

[0067] (4) Transfer: Remove the gel and place the sponge, filter paper, gel, PVDF membrane (pre-soaked in methanol for 15 seconds), filter paper, and sponge in the order specified by the transfer device, ensuring that there are no bubbles between the layers. Place the transfer device in the transfer buffer in an ice bath and transfer the membrane at a constant current of 300 mA for 2 hours.

[0068] (5) Blocking: After the transfer is completed, remove the PVDF membrane and rinse it with TBST buffer three times, 5 minutes each time; then place the PVDF membrane in 5% skim milk powder solution and block it by slowly shaking on a shaker at room temperature for 2 hours;

[0069] (6) Primary antibody incubation: After blocking, rinse the PVDF membrane three times with TBST buffer, 5 min each time; add diluted primary antibodies against type I collagen (1:1000) and α-SMA protein (1:1000), respectively, and incubate overnight on a shaker at 4°C. At the same time, set the internal reference β-actin primary antibody (1:5000) for incubation, and the operation is the same as above;

[0070] (7) Secondary antibody incubation: After the primary antibody incubation, the PVDF membrane was rinsed with TBST buffer three times, 10 min each time; HRP-labeled corresponding secondary antibody (1:5000) was added and incubated at room temperature with slow shaking on a shaker for 1.5 h;

[0071] (8) Color development and imaging: after the incubation of the secondary antibody, the PVDF membrane was washed with TBST buffer for 3 times, 10 min each time; ECL chemiluminescence reagent A and B were mixed in a ratio of 1:1, evenly dropped on the PVDF membrane, and exposed and imaged in a chemiluminescence imaging system, and the bands were recorded;

[0072] (9) Gray scale analysis: the gray value of the imaged bands was analyzed using ImageJ software, and the ratio of the gray value of the collagen type I protein and alpha-SMA protein bands to the gray value of the internal reference beta-actin band was calculated as the relative expression of each protein;

[0073] Table 3 Detection results of fibrosis-related proteins

[0074]

[0075]

[0076] As can be seen from Table 3, the expression of collagen type I protein and alpha-SMA protein in the ventricular tissue of DM group rats was significantly increased, indicating that diabetes caused significant ventricular fibrosis; each administration group can inhibit the fibrosis process to a certain extent, and the inhibition effect of Example 1 group is the best, indicating that spermidine, coenzyme Q10, panax notoginseng saponins and resveratrol have a stronger effect in reducing ventricular fibrosis when the four components act synergistically.

[0077] Experimental Example 3 Determination of myocardial cell apoptosis

[0078] The proportion of apoptotic cells in myocardial tissue was detected by TUNEL staining method, and the expression level of apoptosis-related proteins (Bax, Bcl-2, Cleaved-Caspase-3) was determined by Western blot method, and the detection results are shown in Table 4;

[0079] TUNEL staining method:

[0080] The fixed tissue was dehydrated and paraffin-embedded in turn, and 5 μm heart sections were prepared using a microtome; the sections were dewaxed by xylene bath and rehydrated by a series of decreasing concentration of ethanol; cell apoptosis detection was performed using a TUNEL detection kit (purchased from Shanghai Hengfei Biological Technology Co., Ltd.);

[0081] Western blot method for determining apoptosis-related proteins: refer to Experimental Example 2;

[0082] Table 4 Detection results of the proportion of apoptotic cells and apoptosis-related proteins

[0083]

[0084] As can be seen from Table 4, the cardiomyocyte apoptosis rate in the DM group was significantly increased, the pro-apoptotic proteins (Bax, Cleaved-Caspase-3) were highly expressed, and the anti-apoptotic protein (Bcl-2) was lowly expressed, indicating that cardiomyocyte apoptosis is an important pathological feature of diabetic cardiomyopathy; the compositions of Example 1, Comparative Example 1, and Comparative Example 2 can all inhibit cardiomyocyte apoptosis, but the effect of Example 1 is better (the apoptosis rate is more significantly reduced, and the protein expression is more obviously improved), indicating that when the four ingredients of spermidine, coenzyme Q10, notoginseng saponins, and resveratrol act synergistically, the inhibitory effect on cardiomyocyte apoptosis is stronger.

[0085] Experimental Example 4 Detection of inflammatory factors

[0086] Myocardial tissue homogenates (prepared as in Experimental Example 2) were obtained from rats in each group (Ctrl group, DM group, Example 1 group, Comparative Example 1 group, and Comparative Example 2 group) in Experimental Example 1 and centrifuged at 12,000 rpm for 10 min at 4°C. The supernatant was stored at -80°C for later use. Before testing, the samples were removed, thawed at room temperature, and centrifuged again (as before). The supernatant was used for testing.

[0087] The concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) in myocardial tissue homogenate were detected using ELISA kits (R&D Systems kits). The test results are shown in Table 5.

[0088] Table 5 Detection results of inflammatory factors

[0089] Group TNF-α (pg / ml) IL-6 (pg / ml) MCP-1 (pg / ml) Control group 45.2±5.3 32.6±4.1 68.5±7.2 DM group 216.8±12.5 185.3±10.2 256.7±15.8 Example 1 group 98.5±8.6 76.2±6.5 125.3±9.4 Comparative Example 1 group 152.6±10.1 128.5±8.3 198.6±12.1 Comparative Example 2 group 135.4±9.2 105.7±7.6 172.5±10.8

[0090] As can be seen from Table 5, the concentrations of the three inflammatory factors in Example 1 group were significantly lower than those in the DM group (P<0.01), and the decrease was greater than that in the Comparative Example 1 group and the Comparative Example 2 group (P<0.05); the inflammation inhibition effects of the Comparative Example 1 group (lacking spermidine) and the Comparative Example 2 group (lacking resveratrol) were weaker, indicating that spermidine and resveratrol play a synergistic role in inhibiting myocardial inflammation.

[0091] Example 4 A tablet for preventing and / or treating diabetic cardiomyopathy (100 tablets)

[0092] Including spermidine 0.05g (purity ≥98%), coenzyme Q10 1.5g (purity ≥99%), notoginseng saponin 1g (purity ≥98%), resveratrol 0.02g (purity ≥98%), microcrystalline cellulose 3g, hypromellose 0.2g, sodium starch glycolate 0.25g and magnesium stearate 0.08g.

[0093] Example 5 A sustained-release capsule for preventing and / or treating diabetic cardiomyopathy (100 capsules)

[0094] Including spermidine 0.03g (purity ≥98%), coenzyme Q10 1.8g (purity ≥99%), notoginseng saponin 0.5g (purity ≥98%), resveratrol 0.01g (purity ≥98%), mannitol 2g, povidone 0.15g, ethyl cellulose 0.5g, and talc 0.1g.

[0095] As can be seen from the above embodiments, the present invention provides a composition for preventing and / or treating diabetic cardiomyopathy and its application. The composition, through the scientific combination of four active ingredients, namely spermidine, coenzyme Q, notoginseng saponins and resveratrol, exhibits significant synergistic effects and multiple protection mechanisms in the treatment of diabetic cardiomyopathy; and the composition has high biosafety characteristics, is easily absorbed and metabolized by the human body, and has few side effects.

[0096] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A composition for preventing and / or treating diabetic cardiomyopathy, characterized in that: It includes the following ingredients: spermidine, coenzyme Q, notoginseng saponin and resveratrol; The mass ratio of spermidine, coenzyme Q, notoginseng saponin and resveratrol is 0.3-1:6-20:3-15:0.1-0.

5.

2. The composition according to claim 1, characterized in that The mass ratio of spermidine, coenzyme Q, notoginseng saponin and resveratrol is 0.5-0.8:10-16:6-12:0.2-0.

4.

3. Use of the composition according to claim 1 or 2 in the preparation of a preparation for preventing and / or treating diabetic cardiomyopathy.

4. A preparation for preventing and / or treating diabetic cardiomyopathy, characterized in that: The invention comprises the composition according to claim 1 or 2 and pharmaceutically acceptable excipients.

5. The preparation according to claim 4, characterized in that The auxiliary materials include one or more of fillers, binders, disintegrants, lubricants, solvents and sustained-release materials.

6. The preparation according to claim 5, characterized in that The dosage form of the preparation is tablet, capsule, granule, injection or oral solution.

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